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MDRL lncRNA regulates the processing of miR-484 primary transcript by targeting miR-361.

Wang K, Sun T, Li N, Wang Y, Wang JX, Zhou LY, Long B, Liu CY, Liu F, Li PF - PLoS Genet. (2014)

Bottom Line: The results showed that miR-361 that predominantly located in nucleus can directly bind to primary transcript of miR-484 (pri-miR-484) and prevent its processing by Drosha into pre-miR-484. miR-361 is able to regulate mitochondrial fission and apoptosis by regulating miR-484 levels.Our present study reveals a novel regulating model of mitochondrial fission program which is composed of MDRL, miR-361 and miR-484.Our work not only expands the function of the lncRNA pathway in gene regulation but also establishes a new mechanism for controlling miRNA expression.

View Article: PubMed Central - PubMed

Affiliation: Division of Cardiovascular Research, State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.

ABSTRACT
Long noncoding RNAs (lncRNAs) are emerging as new players in gene regulation, but whether lncRNAs operate in the processing of miRNA primary transcript is unclear. Also, whether lncRNAs are involved in the regulation of the mitochondrial network remains to be elucidated. Here, we report that a long noncoding RNA, named mitochondrial dynamic related lncRNA (MDRL), affects the processing of miR-484 primary transcript in nucleus and regulates the mitochondrial network by targeting miR-361 and miR-484. The results showed that miR-361 that predominantly located in nucleus can directly bind to primary transcript of miR-484 (pri-miR-484) and prevent its processing by Drosha into pre-miR-484. miR-361 is able to regulate mitochondrial fission and apoptosis by regulating miR-484 levels. In exploring the underlying molecular mechanism by which miR-361 is regulated, we identified MDRL and demonstrated that it could directly bind to miR-361 and downregulate its expression levels, which promotes the processing of pri-miR-484. MDRL inhibits mitochondrial fission and apoptosis by downregulating miR-361, which in turn relieves inhibition of miR-484 processing by miR-361. Our present study reveals a novel regulating model of mitochondrial fission program which is composed of MDRL, miR-361 and miR-484. Our work not only expands the function of the lncRNA pathway in gene regulation but also establishes a new mechanism for controlling miRNA expression.

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MDRL regulates mitochondrial fission and apoptosis through targeting miR-361 and miR-484.A. Knockdown of MDRL reduces the expression levels of miR-484. Cardiomyocytes were infected with adenoviral MDRL-siRNA or MDRL-sc. 24 h after infection miR-484 levels were analyzed by northern blot. B. Enforced expression of MDRL induces the increase of miR-484 levels. Cardiomyocytes were infected with adenoviral MDRL or β-gal. 24 h after infection miR-484 levels were analyzed by northern blot. C. MDRL reduces the inhibitory effect of miR-361 on miR-484 expression. Cardiomyocytes were infected with adenoviral miR-361, MDRL or β-gal. miR-484 expression levels were analyzed by northern blot. D. MDRL inhibits mitochondrial fission induced by A/R. Cardiomyocytes were infected with adenoviral MDRL or β-gal, and were exposed to A/R. The cells were stained with MitoTracker Green (left panel), Bar = 20 µm. The cells with fragmented mitochondria were counted (right panel). *p<0.05 vs A/R alone. E. MDRL inhibits apoptosis induced by A/R. Cardiomyocytes were infected with adenoviral MDRL or β-gal, then were exposed to A/R. Apoptosis was assessed by TUNEL assay. *p<0.05 vs A/R alone. F. MDRL could inhibit mitochondrial fission and apoptosis upon I/R. Intracoronary delivery of adenoviral constructs of MDRL or β-gal to the hearts was described in the section of Materials and Methods. Mice were subjected to 45 min ischemia and 3 h reperfusion. Counting of fragmented mitochondria and apoptosis were shown. *p<0.05 versus I/R alone. G. MDRL attenuates myocardial infarction upon I/R. Mice were treated as described in (F). The infarct sizes were shown. *p<0.05. Bar = 2 mm. H. Knockdown of miR-484 attenuates the inhibitory effect of MDRL on mitochondria fission and apoptosis. Cardiomyocytes were infected with the adenoviral MDRL, miR-484 antagomir or antagomir-NC, and then treated with A/R. Mitochondria fission and apoptosis were analyzed. *p<0.05.
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pgen-1004467-g006: MDRL regulates mitochondrial fission and apoptosis through targeting miR-361 and miR-484.A. Knockdown of MDRL reduces the expression levels of miR-484. Cardiomyocytes were infected with adenoviral MDRL-siRNA or MDRL-sc. 24 h after infection miR-484 levels were analyzed by northern blot. B. Enforced expression of MDRL induces the increase of miR-484 levels. Cardiomyocytes were infected with adenoviral MDRL or β-gal. 24 h after infection miR-484 levels were analyzed by northern blot. C. MDRL reduces the inhibitory effect of miR-361 on miR-484 expression. Cardiomyocytes were infected with adenoviral miR-361, MDRL or β-gal. miR-484 expression levels were analyzed by northern blot. D. MDRL inhibits mitochondrial fission induced by A/R. Cardiomyocytes were infected with adenoviral MDRL or β-gal, and were exposed to A/R. The cells were stained with MitoTracker Green (left panel), Bar = 20 µm. The cells with fragmented mitochondria were counted (right panel). *p<0.05 vs A/R alone. E. MDRL inhibits apoptosis induced by A/R. Cardiomyocytes were infected with adenoviral MDRL or β-gal, then were exposed to A/R. Apoptosis was assessed by TUNEL assay. *p<0.05 vs A/R alone. F. MDRL could inhibit mitochondrial fission and apoptosis upon I/R. Intracoronary delivery of adenoviral constructs of MDRL or β-gal to the hearts was described in the section of Materials and Methods. Mice were subjected to 45 min ischemia and 3 h reperfusion. Counting of fragmented mitochondria and apoptosis were shown. *p<0.05 versus I/R alone. G. MDRL attenuates myocardial infarction upon I/R. Mice were treated as described in (F). The infarct sizes were shown. *p<0.05. Bar = 2 mm. H. Knockdown of miR-484 attenuates the inhibitory effect of MDRL on mitochondria fission and apoptosis. Cardiomyocytes were infected with the adenoviral MDRL, miR-484 antagomir or antagomir-NC, and then treated with A/R. Mitochondria fission and apoptosis were analyzed. *p<0.05.

Mentions: Our present results have demonstrated that MDRL could promote the processing of pri-miR-484 by Drosha. Thus, we tested whether MDRL is able to regulate mature miR-484 levels. Knockdown of MDRL reduced miR-484 levels (Figure 6A), while overexpression of MDRL resulted in up-regulation of miR-484 expression (Figure 6B). And MDRL counteracted the effect of miR-361 on miR-484 expression (Figure 6C). Our previous report has demonstrated that Fis1 is a downstream target of miR-484. The current data showed that MDRL could regulate Fis1 expression by miR-484 (Figure S5A). These results indicated that MDRL may act as endogenous sponge “antagomir” of miR-361 to regulate the processing of pri-miR-484 and expression of miR-484.


MDRL lncRNA regulates the processing of miR-484 primary transcript by targeting miR-361.

Wang K, Sun T, Li N, Wang Y, Wang JX, Zhou LY, Long B, Liu CY, Liu F, Li PF - PLoS Genet. (2014)

MDRL regulates mitochondrial fission and apoptosis through targeting miR-361 and miR-484.A. Knockdown of MDRL reduces the expression levels of miR-484. Cardiomyocytes were infected with adenoviral MDRL-siRNA or MDRL-sc. 24 h after infection miR-484 levels were analyzed by northern blot. B. Enforced expression of MDRL induces the increase of miR-484 levels. Cardiomyocytes were infected with adenoviral MDRL or β-gal. 24 h after infection miR-484 levels were analyzed by northern blot. C. MDRL reduces the inhibitory effect of miR-361 on miR-484 expression. Cardiomyocytes were infected with adenoviral miR-361, MDRL or β-gal. miR-484 expression levels were analyzed by northern blot. D. MDRL inhibits mitochondrial fission induced by A/R. Cardiomyocytes were infected with adenoviral MDRL or β-gal, and were exposed to A/R. The cells were stained with MitoTracker Green (left panel), Bar = 20 µm. The cells with fragmented mitochondria were counted (right panel). *p<0.05 vs A/R alone. E. MDRL inhibits apoptosis induced by A/R. Cardiomyocytes were infected with adenoviral MDRL or β-gal, then were exposed to A/R. Apoptosis was assessed by TUNEL assay. *p<0.05 vs A/R alone. F. MDRL could inhibit mitochondrial fission and apoptosis upon I/R. Intracoronary delivery of adenoviral constructs of MDRL or β-gal to the hearts was described in the section of Materials and Methods. Mice were subjected to 45 min ischemia and 3 h reperfusion. Counting of fragmented mitochondria and apoptosis were shown. *p<0.05 versus I/R alone. G. MDRL attenuates myocardial infarction upon I/R. Mice were treated as described in (F). The infarct sizes were shown. *p<0.05. Bar = 2 mm. H. Knockdown of miR-484 attenuates the inhibitory effect of MDRL on mitochondria fission and apoptosis. Cardiomyocytes were infected with the adenoviral MDRL, miR-484 antagomir or antagomir-NC, and then treated with A/R. Mitochondria fission and apoptosis were analyzed. *p<0.05.
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Related In: Results  -  Collection

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pgen-1004467-g006: MDRL regulates mitochondrial fission and apoptosis through targeting miR-361 and miR-484.A. Knockdown of MDRL reduces the expression levels of miR-484. Cardiomyocytes were infected with adenoviral MDRL-siRNA or MDRL-sc. 24 h after infection miR-484 levels were analyzed by northern blot. B. Enforced expression of MDRL induces the increase of miR-484 levels. Cardiomyocytes were infected with adenoviral MDRL or β-gal. 24 h after infection miR-484 levels were analyzed by northern blot. C. MDRL reduces the inhibitory effect of miR-361 on miR-484 expression. Cardiomyocytes were infected with adenoviral miR-361, MDRL or β-gal. miR-484 expression levels were analyzed by northern blot. D. MDRL inhibits mitochondrial fission induced by A/R. Cardiomyocytes were infected with adenoviral MDRL or β-gal, and were exposed to A/R. The cells were stained with MitoTracker Green (left panel), Bar = 20 µm. The cells with fragmented mitochondria were counted (right panel). *p<0.05 vs A/R alone. E. MDRL inhibits apoptosis induced by A/R. Cardiomyocytes were infected with adenoviral MDRL or β-gal, then were exposed to A/R. Apoptosis was assessed by TUNEL assay. *p<0.05 vs A/R alone. F. MDRL could inhibit mitochondrial fission and apoptosis upon I/R. Intracoronary delivery of adenoviral constructs of MDRL or β-gal to the hearts was described in the section of Materials and Methods. Mice were subjected to 45 min ischemia and 3 h reperfusion. Counting of fragmented mitochondria and apoptosis were shown. *p<0.05 versus I/R alone. G. MDRL attenuates myocardial infarction upon I/R. Mice were treated as described in (F). The infarct sizes were shown. *p<0.05. Bar = 2 mm. H. Knockdown of miR-484 attenuates the inhibitory effect of MDRL on mitochondria fission and apoptosis. Cardiomyocytes were infected with the adenoviral MDRL, miR-484 antagomir or antagomir-NC, and then treated with A/R. Mitochondria fission and apoptosis were analyzed. *p<0.05.
Mentions: Our present results have demonstrated that MDRL could promote the processing of pri-miR-484 by Drosha. Thus, we tested whether MDRL is able to regulate mature miR-484 levels. Knockdown of MDRL reduced miR-484 levels (Figure 6A), while overexpression of MDRL resulted in up-regulation of miR-484 expression (Figure 6B). And MDRL counteracted the effect of miR-361 on miR-484 expression (Figure 6C). Our previous report has demonstrated that Fis1 is a downstream target of miR-484. The current data showed that MDRL could regulate Fis1 expression by miR-484 (Figure S5A). These results indicated that MDRL may act as endogenous sponge “antagomir” of miR-361 to regulate the processing of pri-miR-484 and expression of miR-484.

Bottom Line: The results showed that miR-361 that predominantly located in nucleus can directly bind to primary transcript of miR-484 (pri-miR-484) and prevent its processing by Drosha into pre-miR-484. miR-361 is able to regulate mitochondrial fission and apoptosis by regulating miR-484 levels.Our present study reveals a novel regulating model of mitochondrial fission program which is composed of MDRL, miR-361 and miR-484.Our work not only expands the function of the lncRNA pathway in gene regulation but also establishes a new mechanism for controlling miRNA expression.

View Article: PubMed Central - PubMed

Affiliation: Division of Cardiovascular Research, State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.

ABSTRACT
Long noncoding RNAs (lncRNAs) are emerging as new players in gene regulation, but whether lncRNAs operate in the processing of miRNA primary transcript is unclear. Also, whether lncRNAs are involved in the regulation of the mitochondrial network remains to be elucidated. Here, we report that a long noncoding RNA, named mitochondrial dynamic related lncRNA (MDRL), affects the processing of miR-484 primary transcript in nucleus and regulates the mitochondrial network by targeting miR-361 and miR-484. The results showed that miR-361 that predominantly located in nucleus can directly bind to primary transcript of miR-484 (pri-miR-484) and prevent its processing by Drosha into pre-miR-484. miR-361 is able to regulate mitochondrial fission and apoptosis by regulating miR-484 levels. In exploring the underlying molecular mechanism by which miR-361 is regulated, we identified MDRL and demonstrated that it could directly bind to miR-361 and downregulate its expression levels, which promotes the processing of pri-miR-484. MDRL inhibits mitochondrial fission and apoptosis by downregulating miR-361, which in turn relieves inhibition of miR-484 processing by miR-361. Our present study reveals a novel regulating model of mitochondrial fission program which is composed of MDRL, miR-361 and miR-484. Our work not only expands the function of the lncRNA pathway in gene regulation but also establishes a new mechanism for controlling miRNA expression.

Show MeSH
Related in: MedlinePlus